Molecular Cloning of a cDNA Coping for Copper/Zinc Superoxide Dismutase from Zebrafish and Its Expression in Escherichia coli

Chuian Fu Ken, Jei Fu Shaw, Jen Leih Wu, Chi Tsai Lin

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

A full-length complementary DNA (cDNA) clone encoding a putative copper/zinc Superoxide dismutase (Cu/Zn-SOD) was amplified by a polymerase chain reaction (PCR)-based technique from cDNA synthesized from zebrafish (Danio rerio) mRNA. Nucleotide sequence analysis of this cDNA clone revealed that it comprised a complete open reading frame coding for 154 amino acid residues. The deduced amino acid sequence showed higher identity (73.5-74.3%) with swordfish and shark Cu/Zn-SOD than with Cu/Zn-SOD from mammals (69.6-70.9%) and plants (55.8-56.2%). The ammo acid residues required for coordinating copper and zinc are conserved, as they are present in all reported Cu/Zn-SOD sequences. It lacks a targeting sequence, which suggests that the zebrafish cDNA clone encodes a cytosolic Cu/Zn-SOD. Furthermore, the coding region of Cu/Zn-SOD from zebrafish was introduced into an expression vector, pET-23a(+)-thioredoxin, and transformed into Escherichia coli AD494(DE3)pLysS. A predominant achromatic zone was detected by activity staining of native PAGE, and the expression pattern was shown by Coomassie blue staining of SDS-PAGE. This indicates that the Cu/Zn-SOD cDNA clone can be expressed in E. coli.

Original languageEnglish
Pages (from-to)2863-2867
Number of pages5
JournalJournal of Agricultural and Food Chemistry
Volume46
Issue number7
Publication statusPublished - 1998 Jul 1

Fingerprint

complementary DNA
Cloning
Molecular Cloning
Zebrafish
Danio rerio
Escherichia coli
Superoxide Dismutase
Zinc
Copper
molecular cloning
superoxide dismutase
Complementary DNA
copper
zinc
Clone Cells
clones
Staining and Labeling
Native Polyacrylamide Gel Electrophoresis
Amino Acids
Sharks

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Agricultural and Biological Sciences(all)

Cite this

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title = "Molecular Cloning of a cDNA Coping for Copper/Zinc Superoxide Dismutase from Zebrafish and Its Expression in Escherichia coli",
abstract = "A full-length complementary DNA (cDNA) clone encoding a putative copper/zinc Superoxide dismutase (Cu/Zn-SOD) was amplified by a polymerase chain reaction (PCR)-based technique from cDNA synthesized from zebrafish (Danio rerio) mRNA. Nucleotide sequence analysis of this cDNA clone revealed that it comprised a complete open reading frame coding for 154 amino acid residues. The deduced amino acid sequence showed higher identity (73.5-74.3{\%}) with swordfish and shark Cu/Zn-SOD than with Cu/Zn-SOD from mammals (69.6-70.9{\%}) and plants (55.8-56.2{\%}). The ammo acid residues required for coordinating copper and zinc are conserved, as they are present in all reported Cu/Zn-SOD sequences. It lacks a targeting sequence, which suggests that the zebrafish cDNA clone encodes a cytosolic Cu/Zn-SOD. Furthermore, the coding region of Cu/Zn-SOD from zebrafish was introduced into an expression vector, pET-23a(+)-thioredoxin, and transformed into Escherichia coli AD494(DE3)pLysS. A predominant achromatic zone was detected by activity staining of native PAGE, and the expression pattern was shown by Coomassie blue staining of SDS-PAGE. This indicates that the Cu/Zn-SOD cDNA clone can be expressed in E. coli.",
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Molecular Cloning of a cDNA Coping for Copper/Zinc Superoxide Dismutase from Zebrafish and Its Expression in Escherichia coli. / Ken, Chuian Fu; Shaw, Jei Fu; Wu, Jen Leih; Lin, Chi Tsai.

In: Journal of Agricultural and Food Chemistry, Vol. 46, No. 7, 01.07.1998, p. 2863-2867.

Research output: Contribution to journalArticle

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